Evaluation of quality parameters in thawed dendritic cell vaccine

Abstract

Background & Aim Manufacturing of Dendritic Cell (DC) vaccine for cancer, a type of Advanced Therapy Medicinal Products, is regulated by cGMP rules. Our experience of DC vaccine preparation started in 2001 and was tested in many clinical trials. Methods, Results & Conclusion DC vaccines were prepared from adherent fraction of Peripheral Blood Mononuclear Cells obtained from leukapheresis, by differentiation and maturation with standard cytokine cocktail after pulsing with autologous tumor homogenate. Aliquots of DC vaccine were frozen by automatic freezing in autologous plasma with 10% DMSO and stored in nitrogen vapors until their use. These aliquots were thawed and resuspended in sterile saline at a final concentration of 5 × 106 cells/ml, packed in two syringes for a total of 10 million that represent the thawed final product. To ensure the maintaining of compliance with release parameters of the thawed final product, three representative batches, stored at 2-8°C for 8 hours, were analyzed using the following tests: Mycoplasma Detection (Method EP. 2.6.7), Bacterial Endotoxin levels (Method EP 2.6.14), Sterility (Method EP 2.6.27) Viable Cell Count (Method EP 2.7.29) Phenotype analysis (internal method) Potency test (ELISPOT COSTIM assay was performed to establish the potency of DCs. We modified the COSTIM assay, which uses IFN-gamma ELISPOT as a read-out system. In this assay DCs provide responder allogeneic T cells, sub-optimally pre stimulated with OKT3, with the costimulatory signals required for full activation). The results and the acceptance criteria are listed in Table 1. Manufacturing of Dendritic Cell (DC) vaccine for cancer, a type of Advanced Therapy Medicinal Products, is regulated by cGMP rules. Our experience of DC vaccine preparation started in 2001 and was tested in many clinical trials. DC vaccines were prepared from adherent fraction of Peripheral Blood Mononuclear Cells obtained from leukapheresis, by differentiation and maturation with standard cytokine cocktail after pulsing with autologous tumor homogenate. Aliquots of DC vaccine were frozen by automatic freezing in autologous plasma with 10% DMSO and stored in nitrogen vapors until their use.